1、BRITISH STANDARDBS EN ISO 15303:2008Animal and vegetable fats and oils Detection and identification of a volatile organic contaminant by GC/MSICS 67.200.10NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWLicensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Un
2、controlled Copy, (c) BSINational forewordThis British Standard is the UK implementation of EN ISO 15303:2008. It is identical with ISO 15303:2001. It supersedes BS ISO 15303:2001, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee AW/11, Animal and vegeta
3、ble fats and oils.A list of organizations represented on this committee can be obtained on request to its secretary. This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application.Compliance with a British Standard cannot co
4、nfer immunity from legal obligations.BS EN ISO 15303:2008This British Standard was published under the authority of the Standards Policy and Strategy Committee on 19 October 2001 BSI 2009ISBN 978 0 580 63596 0Amendments/corrigenda issued since publication Date Comments 30 September 2009 This corrige
5、ndum renumbers BS ISO 15303:2001 as BS EN ISO 15303:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSIEUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 15303November 2008ICS 67.200.10English VersionAnimal and vegetable fats and oils - Detectio
6、n and identificationof a volatile organic contaminant by GC/MS (ISO 15303:2001)Corps gras dorigines animale et vgtale - Dtection etidentification dun contaminant organique volatil parCPG/SM (ISO 15303:2001)Tierische und pflanzliche Fette und le - Nachweis undIdentifizierung einer flchtigen organisch
7、en Verunreinigungmittels GC/MS (ISO 15303:2001)This European Standard was approved by CEN on 23 October 2008.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alterat
8、ion. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the CEN Management Centre or to any CEN member.This European Standard exists in three official versions (English, French, German). A version in any other language made by translat
9、ionunder the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,France, Germany, Gree
10、ce, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Ce
11、ntre: rue de Stassart, 36 B-1050 Brussels 2008 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 15303:2008: ELicensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSIForeword The text of I
12、SO 15303:2001 has been prepared by Technical Committee ISO/TC 34 “Agricultural food products” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 15303:2008 by Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products
13、 - Methods of sampling and analysis” the secretariat of which is held by AFNOR. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by May 2009, and conflicting national standards shall be withdrawn at t
14、he latest by May 2009. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN/CENELEC Internal Regulations, the national
15、 standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Pola
16、nd, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 15303:2001 has been approved by CEN as a EN ISO 15303:2008 without any modification. ii BS EN ISO 15303:2008EN ISO 15303:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA ST
17、ANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSIiiiContents Page1 Scope 12 Normative reference 13 Principle14 Reagents.25 Apparatus .26 Procedure .36.1 Preparation of standard and blank 36.2 Preparation of internal standards 36.3 Preparation of analytical sample36.4 Determination by GC/MS analy
18、sis .36.5 Identification.47 Calculation and expression of results.47.1 Calculation47.2 Expression of results 58 Precision.58.1 Interlaboratory tests 58.2 Repeatability.58.3 Reproducibility.5Annex A (informative) Results of interlaboratory tests.6Bibliography10BS EN ISO 15303:2008EN ISO 15303:2008Lic
19、ensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSIblankLicensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSIINTENRATIONAL TSANDADR1Animal and vegetable fats and oils Detection and identificationof a volatile
20、organic contaminant by GC/MS1 ScopeThis International Standard specifies a method for the detection and identification of a volatile organic contaminantin edible oils.It is applicable to the identification of volatile industrial chemicals in both crude and refined edible oils that aresuspected of be
21、ing contaminated. It also enables determination of the concentration of the contaminant.This International Standard is not applicable to the determination of the concentration of chemicals that may reactwith the edible oil or with one of its natural components. In these cases, the presence of the co
22、ntaminant maysometimes be established on a qualitative basis. Also, this International Standard is not applicable to non-volatilechemicals.This method has been shown to be applicable for the identification of the following compound classes:Gbe saturated halogenated hydrocarbons;Gbe unsaturated halog
23、enated hydrocarbons;Gbe esters; aldehydes; alcohols; amines; ketones; ethers;Gbe cyclic and aromatic compounds;Gbe nitrogen compounds;Gbe acrylates; etc.The method has been evaluated for concentrations in the range of 1 mg/kg to 10 mg/kg.2 Normative referenceThe following normative document contains
24、 provisions which, through reference in this text, constitute provisions ofthis International Standard. For dated references, subsequent amendments to, or revisions of, any of thesepublications do not apply. However, parties to agreements based on this International Standard are encouraged toinvesti
25、gate the possibility of applying the most recent edition of the normative document indicated below. Forundated references, the latest edition of the normative document referred to applies. Members of ISO and IECmaintain registers of currently valid International Standards.ISO 661:1989, Animal and ve
26、getable fats and oils Preparation of test sample3PrincipleA deuterated reference compound with a GC retention time close to that of the suspected contaminant is added tothe oil at a concentration close to that of the suspected contaminant. A sample of the oil is then introduced into a1BS EN ISO 1530
27、3:2008EN ISO 15303:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSI2 thermal cold-trapping inlet of a GC/MS apparatus. Volatile components evaporate from the oil and are held in thecold trap. The trap is then flash-heated to 160 C and the chemic
28、als released from the trap are swept into theGC/MS for analysis.4 ReagentsUse only reagents of recognized analytical grade, and distilled or demineralized water or water of equivalent purity.4.1 Standard reference compound, corresponding to suspected contaminating compound (99 % pure).4.2 Methanol,
29、Analar grade.4.3 Fully deuterated internal standards of benzene, ethyl benzene or naphthalene (99 % pure).4.4 Helium, chemically pure grade.4.5 Refined, bleached, deodorized (RBD) groundnut oil, or a similar stable vegetable liquid oil, known to befree of industrial chemicals related to the analyte
30、under consideration.5 ApparatusUsual laboratory apparatus and, in particular, the following.5.1 Capillary gas chromatograph.5.2 Mass spectrometer.5.3 Thermal desorption cold-trapping device1).5.4 Capillary column, of length 50 m, methyl polysiloxane with OV101 (or equivalent) stationary phase, 0,5 m
31、film thickness and 0,32 mm internal diameter.5.5 Gas chromatograph/mass spectrometer, operating under the following conditions.a) Thermal desorption cold-trapping temperature programme (at injection):Gbe thermal cold trap oven 160 C;Gbe thermal cold trap below 40 C.The trap is held at a temperature
32、below 40 C for 5 min and is then flash-heated to 160 C. The GCtemperature programme is started simultaneously with the flash-heating of the trap. The trap is held at 160 Cfor 3 min.b) GC temperature programme:Gbe initial temperature 50 Cfor5min;Gbe increase at 7,5 C per min;Gbe final temperature 250
33、 Cfor5min.1) The Chrompack Purge and Trap System is an example of a suitable product available commercially. This information isgiven for the convenience of users of this International Standard and does not constitute an endorsement by ISO of the product.BS EN ISO 15303:2008EN ISO 15303:2008Licensed
34、 Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSI3c) MS conditions:Gbe scan one scan per second or faster;Gbe source 70 eV, 200 C, 100 A, 4 kV.d) Gas flows:Gbe column 1 ml/min helium;Gbe split during cold trapping 5 ml/min.5.6 Vortex mixer.6 Procedure6.1 Pre
35、paration of standard and blank6.1.1 Use a refined, bleached and deodorized vegetable oil e.g. groundnut (4.5) as a carrier and as a blank.6.1.2 Make up the standard (4.1) (i.e. a pure chemical of known composition, corresponding to that of thesuspected contaminant) in a concentrated form (e.g. 100 m
36、g/kg) in the carrier oil (6.1.1). This is the stock solution.Dilute this stock solution with carrier oil to the required concentration (usually in the range 1 mg/kg to 10 mg/kg).The standard solution should be of a similar concentration to the contaminant in the sample.6.1.3 Dilute the suspect oil,
37、if necessary, with the carrier oil (4.5) so that the concentration of the suspectedcontaminant is likely to be in the range 1,0 mg/kg to 10 mg/kg, i.e. the established linear range for quantificationwith this method.6.2 Preparation of internal standardsPrepare a solution of deuterated benzene, ethyl
38、 benzene or naphthalene (4.3) in methanol (4.2) at a concentrationof 0,1 g/l (0,1 g/l) as needed. The internal standard chosen should be the one having the closest GLC retentiontime to that of the analyte.6.3 Preparation of analytical sampleAccurately weigh 1,00 g of the oil under test. Add 5,0 l of
39、 internal standard solution in methanol (4.2), selectedaccording to 6.2. Mix on the vortex mixer (5.6).Repeat with all samples, standard and blank. For best results, allow to stand overnight to allow equilibration.6.4 Determination by GC/MS analysisThe following conditions have been found to be sati
40、sfactory.Pack an empty thermal cold-trapping (TCT) tube with clean glass wool to a length of about 3 cm to 4 cm. Takeapproximately 10 mg of sample and place in the tube. If the observed concentration of the contaminant exceeds10 mg/kg, following analysis of a 10 mg portion of oil, repeat the determi
41、nation with a sample size of 2,5 mg. Forstill larger concentrations, it is recommended that the sample be accurately diluted with the blank oil (6.1.1) until theconcentration range is 1 mg/kg to 10 mg/kg (see 6.1.3).Pre-cool the cold trap to below 20 C, and continue cooling to below 40 C. Fit the tu
42、be containing the oil in theTCT oven and start the TCT and GC/MS programmes.BS EN ISO 15303:2008EN ISO 15303:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSI4 6.5 IdentificationCompare the GC retention time and mass spectrum of the analyte with
43、those of the standard. Identification requiresa good match of both spectra and GC retention times.Quantification is performed by using reconstructed ion chromatograms of selected ions from the analyte and theinternal standard and integrating their areas. This method is used to minimize interference
44、from co-elutants.7 Calculation and expression of results7.1 CalculationThe peak area of the analyte ion divided by the peak area of the internal standard ion is the response ratio R:isKCRCGd7G3dwhereK is a constant;C is the concentration of analyte ion;Cisis the concentration of internal standard.SA
45、MSAMisRKCCG3dGd7whereRSAMis the value of R for the sample;CSAMis the concentration of analyte ion in the sample.Since Cisis a constant by definition:SAM SAMRFCG3dGd7where the factor F isisKFCG3dSimilarly, the value of R for the standard is:STD STDRFCG3dGd7where CSTDis the concentration of analyte io
46、n in the standard.BS EN ISO 15303:2008EN ISO 15303:2008Licensed Copy: Wang Bin, ISO/EXCHANGE CHINA STANDARDS, 24/12/2009 02:18, Uncontrolled Copy, (c) BSI5ThereforeSAM SAMSTD STDCFRCFRGd7G3dGd7SAM STDSAMSTDRCCRGd7G3d7.2 Expression of resultsCarry out duplicate determinations. Report the identity of
47、the contaminant and the mean of the two duplicate valuesobtained for its concentration, provided the repeatability limit in 8.2 is satisfied. Otherwise, repeat the determinationon two further test portions. If this time the difference again exceeds 0,6 mg/kg, take as the result the arithmeticmean of
48、 the four determinations, provided that the maximum difference between the individual results does notexceed 1,0 mg per kilogram of oil.Report the result to one decimal place.8 Precision8.1 Interlaboratory testsDetails of interlaboratory tests on the precision of the method are summarized in annex A
49、. The values derived fromthese interlaboratory tests may not be applicable to concentration ranges and matrices other than those given.8.2 RepeatabilityThe absolute difference between two independent single test results, obtained using the same method on identicaltest material in the same laboratory by the same operator using the same equipment within a short interval of time,will in not more than 5 % of cases be greater than 0,6 mg/kg.8.3 ReproducibilityThe absolute difference between two single test results, obtained using the same method on ide
